Journal: Cell Stem Cell

Article Title: Selective advantage of mutant stem cells in human clonal hematopoiesis is associated with attenuated response to inflammation and aging

doi: 10.1016/j.stem.2024.05.010

Figure Lengend Snippet:

Article Snippet: APC anti-human CD271 (clone ME20.4-1.H4) , Miltenyi , Cat# 130-113-418; RRID: AB_2733363.

Techniques: Recombinant, Reverse Transcription, DNA Library Preparation, DNA Sequencing, RNA Sequencing, Purification, Sequencing, Software, Sterility

Journal:

Article Title: Detergent-insoluble GPI-anchored Proteins Are Apically Sorted in Fischer Rat Thyroid Cells, but Interference with Cholesterol or Sphingolipids Differentially Affects Detergent Insolubility and Apical Sorting

doi:

Figure Lengend Snippet: Steady-state distribution and delivery to the plasma membrane of PLAP and NTR-PLAP in transfected FRT cells. (A) Schemes of PLAP and NTR-PLAP (a fusion protein containing the ectodomain of p75NTR and the C-terminal GPI attachment signal of PLAP). (B) FRT cells stably expressing PLAP or NTR-PLAP were grown to confluence and pulsed overnight with [35S]met–cys or [35S]cys, respectively. After surface biotinylation from the apical (Ap) and basolateral (BL) domains, cells were lysed, immunoprecipitated against specific antibodies, and reprecipitated against streptavidin. (C) FRT cells were labeled for 20 min with [35S]met–cys or [35S]cys. At different times of the chase, surface proteins were biotinylated from the apical (Ap) or the basolateral (BL) side. Cells were then lysed and immunoprecipitated with antibodies against PLAP or NTR and with streptavidin beads. Samples were run on 10% SDS-PAGE and detected by fluorography. PLAP and NTR-PLAP are directly sorted to the apical surface in FRT cells, whereas a small percentage of the two proteins is delivered with similar kinetics to the basolateral domain.

Article Snippet: The polyclonal antibody against PLAP was from Rockland (Gilbertsville, PA), and anti-p75NTR mAb was a gift from Andrè Le Bivic (Institut de Biologie du Développement de Marseille, Marseille, France).

Techniques: Transfection, Stable Transfection, Expressing, Immunoprecipitation, Labeling, SDS Page

Journal:

Article Title: Detergent-insoluble GPI-anchored Proteins Are Apically Sorted in Fischer Rat Thyroid Cells, but Interference with Cholesterol or Sphingolipids Differentially Affects Detergent Insolubility and Apical Sorting

doi:

Figure Lengend Snippet: Pulse-chase analysis and purification on sucrose gradients of transmembrane forms of PLAP and p75NTR in transfected FRT cells. (A) Schemes of p75NTR and PLAP-PS321 (a fusion protein containing the ectodomain of PLAP and the transmembrane and the cytoplasmic domain of the PS321 NTR mutant). (B) Pulse-chase and TX-100 extraction of p75NTR and PLAP-PS321. FRT cells expressing p75NTR and PLAP-PS321 were labeled with [35S]cys or [35S]met–cys, respectively, and subsequently treated as described in Figure ​Figure2A.2A. (C) Purification of p75NTR- and PLAP-PS321–enriched fractions on sucrose density gradients. Labeled proteins of FRT cells expressing p75NTR and PLAP-PS321 were solubilized in TX-100 and purified as described in Figure ​Figure2B.2B. Both transmembrane proteins are soluble in TX-100 and do not float on sucrose density gradients.

Article Snippet: The polyclonal antibody against PLAP was from Rockland (Gilbertsville, PA), and anti-p75NTR mAb was a gift from Andrè Le Bivic (Institut de Biologie du Développement de Marseille, Marseille, France).

Techniques: Pulse Chase, Purification, Transfection, Mutagenesis, Expressing, Labeling

Journal:

Article Title: Detergent-insoluble GPI-anchored Proteins Are Apically Sorted in Fischer Rat Thyroid Cells, but Interference with Cholesterol or Sphingolipids Differentially Affects Detergent Insolubility and Apical Sorting

doi:

Figure Lengend Snippet: Surface expression of GPI-anchored, secretory, and transmembrane proteins after FB1 treatment. (A) FRT cells stably expressing PLAP, NTR-PLAP, or gD1-DAF were grown to confluence in the absence (−) or presence (+) of FB1, as described in MATERIALS AND METHODS. After 30 min of labeling with [35S]met–cys or [35S]cys and a 2-h chase, cells were surface biotinylated from the apical (Ap) or the basolateral (BL) domain, lysed, immunoprecipitated against specific antibodies, and reprecipitated against streptavidin. (B) Amounts of labeled proteins were quantified from three independent experiments and are shown as means ± SD. Hatched bars represent apical proteins, and black bars represent basolateral proteins. Apical PLAP and NTR-PLAP, but not basolateral gD1-DAF, became unpolarized after FB1 treatment. (C) FRT cells expressing PLAP-sec, NTR-sec, p75NTR, or PLAP-PS321 were grown to confluence in the absence (−) or presence (+) of FB1, as described in MATERIALS AND METHODS. After labeling with [35S]met–cys or [35S]cys for 20 min and a chase of 2 h, apical and basolateral media were collected separately and cells were surface biotinylated from apical or basolateral domains. Polarized secretion of the secretory forms was revealed by immunoprecipitation in the apical and basolateral medium (top two panels), whereas biotinylated transmembrane proteins were revealed by double immunoprecipitation with specific antibody and streptavidin beads (bottom two panels). (D) Amounts of labeled proteins were quantified from three independent experiments and are shown as means ± SD. Hatched bars represent apical proteins, and black bars represent basolateral proteins. FB1 does not alter the sorting of transmembrane and secretory proteins.

Article Snippet: The polyclonal antibody against PLAP was from Rockland (Gilbertsville, PA), and anti-p75NTR mAb was a gift from Andrè Le Bivic (Institut de Biologie du Développement de Marseille, Marseille, France).

Techniques: Expressing, Stable Transfection, Labeling, Immunoprecipitation

Journal:

Article Title: Detergent-insoluble GPI-anchored Proteins Are Apically Sorted in Fischer Rat Thyroid Cells, but Interference with Cholesterol or Sphingolipids Differentially Affects Detergent Insolubility and Apical Sorting

doi:

Figure Lengend Snippet: Surface expression of GPI-anchored and transmembrane proteins after cholesterol depletion. (A) FRT cells expressing PLAP, NTR-PLAP, gD1-DAF, p75NTR, or PLAP-PS321 were grown on filters and treated with mevinolin (Mev.; 10 μM) for 48 h and M-β-CD (10 mM) for 60 min. Cells were then pulse labeled for 30 min, chased for 2 h, and biotinylated as usual. (B) Amounts of labeled proteins were quantified in three independent experiments and are shown as means ± SD. Hatched bars represent apical (Ap) proteins, and black bars represent basolateral (BL) proteins. Removal of cholesterol does not alter the surface expression of apical and basolateral GPI-anchored and transmembrane proteins.

Article Snippet: The polyclonal antibody against PLAP was from Rockland (Gilbertsville, PA), and anti-p75NTR mAb was a gift from Andrè Le Bivic (Institut de Biologie du Développement de Marseille, Marseille, France).

Techniques: Expressing, Labeling

Journal: Brain Sciences

Article Title: NGFR Gene and Single Nucleotide Polymorphisms, rs2072446 and rs11466162, Playing Roles in Psychiatric Disorders

doi: 10.3390/brainsci12101372

Figure Lengend Snippet: NGFR SNPs analyzed in this study.

Article Snippet: We used a vector, pCMV6- Ngfr -cDNA, containing a full-length mice Ngfr cDNA clone purchased from OriGene Technologies, Rockville, MD, USA.

Techniques: Variant Assay

Journal: Brain Sciences

Article Title: NGFR Gene and Single Nucleotide Polymorphisms, rs2072446 and rs11466162, Playing Roles in Psychiatric Disorders

doi: 10.3390/brainsci12101372

Figure Lengend Snippet: NGFR shRNA and control sequences.

Article Snippet: We used a vector, pCMV6- Ngfr -cDNA, containing a full-length mice Ngfr cDNA clone purchased from OriGene Technologies, Rockville, MD, USA.

Techniques: shRNA, Control

Journal: Brain Sciences

Article Title: NGFR Gene and Single Nucleotide Polymorphisms, rs2072446 and rs11466162, Playing Roles in Psychiatric Disorders

doi: 10.3390/brainsci12101372

Figure Lengend Snippet: Allele and genotype distribution of NGFR gene polymorphisms between SCZ patients and healthy participants.

Article Snippet: We used a vector, pCMV6- Ngfr -cDNA, containing a full-length mice Ngfr cDNA clone purchased from OriGene Technologies, Rockville, MD, USA.

Techniques: Control

Journal: Brain Sciences

Article Title: NGFR Gene and Single Nucleotide Polymorphisms, rs2072446 and rs11466162, Playing Roles in Psychiatric Disorders

doi: 10.3390/brainsci12101372

Figure Lengend Snippet: Allele and genotype distribution of NGFR gene polymorphisms between MDD patients and healthy participants.

Article Snippet: We used a vector, pCMV6- Ngfr -cDNA, containing a full-length mice Ngfr cDNA clone purchased from OriGene Technologies, Rockville, MD, USA.

Techniques: Control

Journal: Brain Sciences

Article Title: NGFR Gene and Single Nucleotide Polymorphisms, rs2072446 and rs11466162, Playing Roles in Psychiatric Disorders

doi: 10.3390/brainsci12101372

Figure Lengend Snippet: GFP expressed by the transfected cells. The 293T cells were co-transfected with shRNA plasmid and pCAGIG- Ngfr -cDNA.

Article Snippet: We used a vector, pCMV6- Ngfr -cDNA, containing a full-length mice Ngfr cDNA clone purchased from OriGene Technologies, Rockville, MD, USA.

Techniques: Transfection, shRNA, Plasmid Preparation

Journal: Brain Sciences

Article Title: NGFR Gene and Single Nucleotide Polymorphisms, rs2072446 and rs11466162, Playing Roles in Psychiatric Disorders

doi: 10.3390/brainsci12101372

Figure Lengend Snippet: Down-regulation of NGFR in 293T cells. ( A ) Immunoblotting of protein extracted from 293T cells co-transfected with shRNA plasmid and pCAGIG- Ngfr -cDNA. ( B ) One-way ANOVA of the protein levels after normalization to β-actin; data showed as means ± SEM, ** p < 0.01.

Article Snippet: We used a vector, pCMV6- Ngfr -cDNA, containing a full-length mice Ngfr cDNA clone purchased from OriGene Technologies, Rockville, MD, USA.

Techniques: Western Blot, Transfection, shRNA, Plasmid Preparation

Journal: Brain Sciences

Article Title: NGFR Gene and Single Nucleotide Polymorphisms, rs2072446 and rs11466162, Playing Roles in Psychiatric Disorders

doi: 10.3390/brainsci12101372

Figure Lengend Snippet: Down-regulation of NGFR in mouse hippocampus. ( A ) Lenti- NGFR -sh2 or control virus was injected into the dentate gyrus of adult mice. Two weeks after injection, extracts from hippocampus were immunoblotted. ( B ) One-way ANOVA of the protein levels after normalization to β-actin, data showed as means ± SEM, * p < 0.05.

Article Snippet: We used a vector, pCMV6- Ngfr -cDNA, containing a full-length mice Ngfr cDNA clone purchased from OriGene Technologies, Rockville, MD, USA.

Techniques: Control, Virus, Injection

Journal: Transgenic Research

Article Title: Redundancy of p75NTR neurotrophin receptor function in development, growth and fertility in the rat

doi: 10.1007/s11248-024-00395-9

Figure Lengend Snippet: Germline transmission efficiency

Article Snippet: A constitutive p75NTR knock-out rat generated by ZNF-directed in ovo gene editing is commercially available ( https://insights.envigo.com/ ).

Techniques: Transmission Assay, Modification

Journal: Transgenic Research

Article Title: Redundancy of p75NTR neurotrophin receptor function in development, growth and fertility in the rat

doi: 10.1007/s11248-024-00395-9

Figure Lengend Snippet: Viability of P75NTR KO rats

Article Snippet: A constitutive p75NTR knock-out rat generated by ZNF-directed in ovo gene editing is commercially available ( https://insights.envigo.com/ ).

Techniques:

Journal: Transgenic Research

Article Title: Redundancy of p75NTR neurotrophin receptor function in development, growth and fertility in the rat

doi: 10.1007/s11248-024-00395-9

Figure Lengend Snippet: In vivo Cre recombinase-mediated deletion of conditional p75NTR allele. a PCR screening strategy used for detecting Cre recombinase-mediated deletion of exon 2 and discriminating between wild-type, p75NTR Ex2-fl and p75NTR Ex2-∆ alleles. The locations of exons (red boxes), non-exonic genomic DNA (thick black lines), loxP sites (pink arrowheads), FRT sites (yellow arrowheads), forward PCR primers (light and dark green arrowheads) and reverse PCR primers (red arrowheads) are shown. Sizes of expected PCR fragments are highlighted. b PCR analysis of genomic DNA from wild-type rat ESCs (WT), and p75NTR Ex2-∆/∆ (∆/∆) and p75NTR Ex2-l/fl (fl/fl) homozygous rats along with offspring (1 & 2) generated from breeding of a male homozygous p75NTR Ex2-fl/fl (fl/flpar) with a female heterozygous Cre transgenic rat. The upper panel shows amplicons generated using the 5’ forward primer (dark green arrowhead) and reverse primer, demonstrating deletion of Exon 2 in p75NTR Ex2-fl/fl /CRE offspring 1 & 2). The lower panel shows amplicons generated using the 3’ forward primer within the deleted region 5’ to exon 2 (lower panel) and the reverse primer. c PCR amplification of a 233 bp DNA fragment from the samples in 2b using primers specific for the CRE transgene. (Colour figure online)

Article Snippet: A constitutive p75NTR knock-out rat generated by ZNF-directed in ovo gene editing is commercially available ( https://insights.envigo.com/ ).

Techniques: In Vivo, Generated, Transgenic Assay, Amplification

Journal: Transgenic Research

Article Title: Redundancy of p75NTR neurotrophin receptor function in development, growth and fertility in the rat

doi: 10.1007/s11248-024-00395-9

Figure Lengend Snippet: Loss of p75NTR expression in KO rats. a RT-quantitative PCR analysis of p75NTR in wildtype (+ / +), heterozygous (∆/ +) and homozygous (∆/∆) p75NTR Ex2-∆ rat brains (mean ± sd of three technical replicates). b Western blot analysis of p75NTR protein expression in wildtype (+ / +), heterozygous (∆/ +) and homozygous (∆/∆) p75NTR Ex2-∆ rat brains. c Western blot analysis of p75NTR protein expression in wildtype (+ / +), and homozygous (fl/fl) p75NTR Ex2-fl rat brains. The images are composites of P75NTR and βtubulin exposures using the LICOR fluorescent detection system and bands located at 25-37 kDa were detected in the βtubulin channel

Article Snippet: A constitutive p75NTR knock-out rat generated by ZNF-directed in ovo gene editing is commercially available ( https://insights.envigo.com/ ).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Transgenic Research

Article Title: Redundancy of p75NTR neurotrophin receptor function in development, growth and fertility in the rat

doi: 10.1007/s11248-024-00395-9

Figure Lengend Snippet: Fertility of P75NTR KO rats

Article Snippet: A constitutive p75NTR knock-out rat generated by ZNF-directed in ovo gene editing is commercially available ( https://insights.envigo.com/ ).

Techniques:

Journal: Transgenic Research

Article Title: Redundancy of p75NTR neurotrophin receptor function in development, growth and fertility in the rat

doi: 10.1007/s11248-024-00395-9

Figure Lengend Snippet: Histopathological analysis of p75NTR Ex2-∆/∆ and p75NTR Ex2-fl/fl rats. a Representative images of haematoxylin and eosin-stained histological sections from male p75NTR Ex2-fl/fl and p75NTR Ex2-∆/∆ homozygous rats. b Whole, fixed brain weight normalised to body weight, for p75NTR Ex2-fl/fl and p75NTR Ex2-∆/∆ homozygous rats. The statistical analysis was performed using an unpaired two sample t-test with a 5% significance level (mean ± sd of three biological replicates). c Cell density in the cerebellum (left panel), caudoputamen (middle panel) and thalamus (right panel) of p75NTR Ex2-fl/fl and p75NTR Ex2-∆/∆ homozygous rats (mean ± sd of three biological replicates)

Article Snippet: A constitutive p75NTR knock-out rat generated by ZNF-directed in ovo gene editing is commercially available ( https://insights.envigo.com/ ).

Techniques: Staining

Journal: Veterinary pathology

Article Title: p75 Neurotrophin Receptor: A Double-Edged Sword in Pathology and Regeneration of the Central Nervous System.

doi: 10.1177/0300985818781930

Figure Lengend Snippet: Figure 1. Schematic representation of the various interactors of nerve growth factor receptor p75 (p75NTR). The extracellular domain binds neurotrophins as well as their respective proforms (red). Inter- action with other receptors (gray) influences the outcome of p75NTR

Article Snippet: Furthermore, the following antibodies have been tested for immunohistochemistry in canine and murine samples and specifically reacted with p75NTR in our lab: polyclonal p75NTR antibody (extracellular domain; Novus Biologicals NBP1-25971) and polyclonal p75NTR antibody (intracellular domain; Alomone labs ANT-011).

Techniques:

Journal: Veterinary pathology

Article Title: p75 Neurotrophin Receptor: A Double-Edged Sword in Pathology and Regeneration of the Central Nervous System.

doi: 10.1177/0300985818781930

Figure Lengend Snippet: Figure 14. Antigen expression in different Schwann cell stages (sche- matic representation). Markers in the violet box are expressed in immature and nonmyelinating Schwann cells but not in myelinating Schwann cells. GalC (blue) is demonstrated in both myelinating and nonmyelinating Schwann cells. a1b1 integrin (yellow) and markers in the green box are specific for nonmyelinating and myelinating Schwann cells, respectively. GalC, galactocerebrosid; GAP-43, growth- associated protein 43; GFAP, glial fibrillary acidic protein; Krox-20, early growth response protein 2 (EGR2); MAG, myelin-associated protein; MAL, myelin and lymphocyte protein; MBP, myelin basic pro- tein; NCAM, neural cell adhesion molecule; p75NTR, nerve growth factor receptor; PLP, proteolipid protein; PMP-22, peripheral myelin protein 22; P2, peripheral myelin protein 2 (PMP2); Ran-2, rat neural antigen-2. Modified from Mirsky et al and Lempp et al.121,135

Article Snippet: Furthermore, the following antibodies have been tested for immunohistochemistry in canine and murine samples and specifically reacted with p75NTR in our lab: polyclonal p75NTR antibody (extracellular domain; Novus Biologicals NBP1-25971) and polyclonal p75NTR antibody (intracellular domain; Alomone labs ANT-011).

Techniques: Expressing, Modification